Evaluating BioEmu-Generated Kinase Ensembles Reveals Structure Selection as the Virtual Screening Bottleneck.

Abstract

Virtual screening (VS) is an essential tool in drug discovery to prioritize potential drug candidates from vast chemical space. One key challenge limiting its performance is accounting for protein conformational flexibility. While ensemble docking methods have been developed to address this challenge by incorporating multiple protein conformations, these methods often rely on computationally intensive physics-based simulations to sample the relevant conformational space. Generative machine learning models offer a highly promising, scalable, and high-throughput alternative to overcome the limitations of these traditional approaches. We therefore investigate whether conformational ensembles generated by BioEmu, a recently developed generative model, can improve VS performance for kinase targets. Using the DUD-E benchmark data set and a validated AutoDock-GPU protocol, we generated and analyzed nearly 1300 structures across 26 kinases (approximately 50 structures each). BioEmu produces structurally diverse ensembles with substantial performance variation among individual structures. However, ensemble methods employing consensus or best-score selection fail to improve upon, and often degrade, VS performance compared to crystal structure baselines. To investigate the source of this limitation, we quantified the relationship between KinCoRe-based conformational state classification and screening performance. By calculating the coefficient of determination (R2) across the kinase subset, we found that the structural features governing VS performance differ substantially from those defining standard conformational states, with KinCoRe classifications leaving over 84% of performance variance unexplained. This critical gap demonstrates that structural diversity alone is insufficient to guarantee screening success. We show that prospective structure selection, rather than structure generation, represents the primary bottleneck in ensemble-based VS, highlighting an urgent need for novel structural descriptors to identify high-performing conformations.